Recombinant Human Turbo Dicer Enzyme Kit

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1. Recombinant Human Turbo Dicer Enzyme Kit

Genlantis A division of Gene Therapy Systems Inc PRODUCT SUMMARY Cat No 1520002 50 Units Kit Components Content _ Recombinant Turbo Dicer Enzyme 1 tube x 100 ul 0 5 unit ul Dicer Reaction Buffer 1 tube x 50 ul 10 mM ATP 1 tube x 50 ul Dicer Stop Solution 1 tube x 100 ul 5X BSA 1 tube x 50 ul Nuclease free Water 1 tube x 1 ml Shipping and Storage The Recombinant Human Turbo Dicer Enzyme Kit is shipped frozen For maximum stability and long term use immediately store at 20 C upon receipt All components are stable for six months when stored properly 50 mM MgCl Solution 1 tube x 100 ul Description The Recombinant Human Turbo Dicer Enzyme Kit uses an ultra active form of human recombinant dicer enzyme that can cleave more than 95 of dsRNA template into 22 bp siRNAs within 2 hours under optimized reaction conditions Figure 1 This provides sufficient siRNAs in every single species to achieve effective gene silencing Fig 1 Efficient Digestion of Double Stranded RNA with Recombinant Human Turbo Dicer Enzyme I 3 Agarose Gel Lane Sample 1 100 bp marker 2 lug 700 bp dsRNA 3 1ug 700 bp dsRNA 15 hour digestion with 1 unit Dicer Enzyme 4 1ug 700 bp dsRNA 2 hour digestion by 1 unit Turbo Dicer Enzyme Recombinant Human Turbo Dicer Enzyme Kit Page 1 of 2 Recombinant Human Turbo Dicer Enzyme Kit NOTE The Recombinant Human Dicer Enzyme Kit is intended FOR RESEARCH USE ONLY METHODS AND PROCEDURES Preliminary Notes 1 We recommend that you use dsRNA templates of 500 1000 bp in length The dicer enzyme might not digest well if the dsRNA is smaller than 300 bp 2 For optimal results we recommend using the GTS TurboScript T7 RNA Transcription Kit Catalog T510003 to generate double stranded RNA dsRNA dsRNAs generated with TurboScript can be used directly with the Recombinant Turbo Dicer Enzyme Kit without purification however dsRNAs purified using standard lithium chloride or equivalent procedures can give slightly better results Generation of siRNA Using Recombinant Turbo Dicer Enzyme 1 Keep the Dicer Reaction Buffer at room temperature while assembling the reaction The following amounts are for a single 10 ul reaction 3 ul x Nuclease free water x ul dsRNA 1 ug 1 ul 10 mM ATP 1 ul 5X BSA 2 ul 50 mM MgCl 1 ul 10X Dicer Reaction Buffer 2 ul Recombinant Turbo Dicer Enzyme 0 5 unit ul NOTE J Ug of dsRNA control template will yield about 0 5 ug of diced siRNA d siRNA after purification with GTS RNA Purification Columns 1 amp 2 this is sufficient for one or two transfections in 24 well plates using the GeneSilencer siRNA Transfection Reagent For more d sIRNA s scale up the reaction volume proportionally IMPORTANT Avoid using excess Recombinant Dicer Enzyme as it may decrease the amount of d siRNA that can be generated from the digestio		 PDF Manual ENGLISH


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